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Beyond PrP9res) type 1/type 2 dichotomy in Creutzfeldt-Jakob disease.

Uro-Coste E, Cassard H, Simon S, Lugan S, Bilheude JM, Perret-Liaudet A, Ironside JW, Haik S, Basset-Leobon C, Lacroux C, Peoch' K, Streichenberger N, Langeveld J, Head MW, Grassi J, Hauw JJ, Schelcher F, Delisle MB, Andréoletti O - PLoS Pathog. (2008)

Bottom Line: However, since cases of CJD show co-occurrence of type 1 and type 2 PrP(res) in the brain, the basis of this classification system and its relationship to agent strain are under discussion.In 30% of cases, both type 1 and type 2 PrP(res) were identified.Despite this, the other two biochemical assays found that PrP(Sc) from an individual patient demonstrated uniform biochemical properties.

View Article: PubMed Central - PubMed

Affiliation: INSERM U858, Institut de Médecine Moléculaire de Rangueil and Service d'Anatomie Pathologique et Histologie-Cytologie, C.H.U. Rangueil, Toulouse, France.

ABSTRACT
Sporadic Creutzfeldt-Jakob disease (sCJD) cases are currently subclassified according to the methionine/valine polymorphism at codon 129 of the PRNP gene and the proteinase K (PK) digested abnormal prion protein (PrP(res)) identified on Western blotting (type 1 or type 2). These biochemically distinct PrP(res) types have been considered to represent potential distinct prion strains. However, since cases of CJD show co-occurrence of type 1 and type 2 PrP(res) in the brain, the basis of this classification system and its relationship to agent strain are under discussion. Different brain areas from 41 sCJD and 12 iatrogenic CJD (iCJD) cases were investigated, using Western blotting for PrP(res) and two other biochemical assays reflecting the behaviour of the disease-associated form of the prion protein (PrP(Sc)) under variable PK digestion conditions. In 30% of cases, both type 1 and type 2 PrP(res) were identified. Despite this, the other two biochemical assays found that PrP(Sc) from an individual patient demonstrated uniform biochemical properties. Moreover, in sCJD, four distinct biochemical PrP(Sc) subgroups were identified that correlated with the current sCJD clinico-pathological classification. In iCJD, four similar biochemical clusters were observed, but these did not correlate to any particular PRNP 129 polymorphism or western blot PrP(res) pattern. The identification of four different PrP(Sc) biochemical subgroups in sCJD and iCJD, irrespective of the PRNP polymorphism at codon 129 and the PrP(res) isoform provides an alternative biochemical definition of PrP(Sc) diversity and new insight in the perception of Human TSE agents variability.

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PrPsc PK Resistance and Molecular Strain Variations in sCJD and iCJD Brain Samples.Each investigated brain sample was initially characterized by WB using antibody Sha31. Symbol patterns represent type 1 in white, type 1+2 in grey and type 2 in black. (A) Results from PK resistance ELISA carried out on three different brain areas (cerebellum, caudate nucleus and temporal cortex) from a MM1 (open circles), a VV1 (open triangles), a VV2 (inverted filled triangle) and a MM2 (filled squares) sCJD patient. Values obtained are expressed as percentage of signal obtained with the lowest PK concentration (50 µg/mL). (B) Results from CEA strain typing ELISA (one symbol per patient—3 to 5 different areas by patients). PrPSc signal intensity was measured after PK digestion into two different detergent solutions. Normalized A/A′ ratio was calculated for each sample (see Methods section). MM1 and MV1 had a low ratio indicating an absence of alteration of PrPsc PK sensitivity linked to the modification detergent digestion conditions. This ratio was higher in MV2, VV2, MV1+2, and VV1+2, while, in the unique VV1 case, an intermediate ratio was observed. In MM2 patients, the huge ratio indicated a strong increase in PK sensitivity by modification of detergent conditions. (C, D) PK resistance assay in three areas from a (C) VV (triangles) or MM (circles) patient and in (D) a MV (triangles) patient harbouring distinct PrPSc WB type in their different brain areas. Artificial mixtures of MM2/VV1 or MM1/VV2 samples were prepared. All homogenates were first equilibrated by dilution into negative brain homogenate to obtain an equal PrPSc signal in ELISA. (E, F) Mixtures were then tested by Western Blot (200 µg PK digestion—Sha31 anti PrP antibody). (E) Lane 1: MM1 100%; Lane 2: MM1 75%/VV2 25%; Lane 3: MM1 50%/VV2 50%; Lane 4: MM1 25%/VV2 75%; Lane 5: VV2 100%. (F) Lane 1: VV1 100%; Lane 2: VV1 75%/MM2 25%; Lane 3: VV1 50%/MM2 50%; Lane 4: VV1 25%/MM2 75%; Lane 5: MM2 100%. (G, H) Same mixtures were tested in the PK resistance ELISA assay. (G) VV2 100% (filled circles), VV2 75%/MM1 25% (filled triangles), VV2 50%/MM1 50% (filled inverted triangles), VV2 25%/MM1 75% (open triangles), MM1 100% (open circles). (H) MM2 100% (filled circles), MM2 75%/VV1 25% (filled triangles), MM2 50%/VV1 50% (filled inverted triangles), MM2 25%/VV1 75% (open triangles), VV1 100% (open circles).
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ppat-1000029-g003: PrPsc PK Resistance and Molecular Strain Variations in sCJD and iCJD Brain Samples.Each investigated brain sample was initially characterized by WB using antibody Sha31. Symbol patterns represent type 1 in white, type 1+2 in grey and type 2 in black. (A) Results from PK resistance ELISA carried out on three different brain areas (cerebellum, caudate nucleus and temporal cortex) from a MM1 (open circles), a VV1 (open triangles), a VV2 (inverted filled triangle) and a MM2 (filled squares) sCJD patient. Values obtained are expressed as percentage of signal obtained with the lowest PK concentration (50 µg/mL). (B) Results from CEA strain typing ELISA (one symbol per patient—3 to 5 different areas by patients). PrPSc signal intensity was measured after PK digestion into two different detergent solutions. Normalized A/A′ ratio was calculated for each sample (see Methods section). MM1 and MV1 had a low ratio indicating an absence of alteration of PrPsc PK sensitivity linked to the modification detergent digestion conditions. This ratio was higher in MV2, VV2, MV1+2, and VV1+2, while, in the unique VV1 case, an intermediate ratio was observed. In MM2 patients, the huge ratio indicated a strong increase in PK sensitivity by modification of detergent conditions. (C, D) PK resistance assay in three areas from a (C) VV (triangles) or MM (circles) patient and in (D) a MV (triangles) patient harbouring distinct PrPSc WB type in their different brain areas. Artificial mixtures of MM2/VV1 or MM1/VV2 samples were prepared. All homogenates were first equilibrated by dilution into negative brain homogenate to obtain an equal PrPSc signal in ELISA. (E, F) Mixtures were then tested by Western Blot (200 µg PK digestion—Sha31 anti PrP antibody). (E) Lane 1: MM1 100%; Lane 2: MM1 75%/VV2 25%; Lane 3: MM1 50%/VV2 50%; Lane 4: MM1 25%/VV2 75%; Lane 5: VV2 100%. (F) Lane 1: VV1 100%; Lane 2: VV1 75%/MM2 25%; Lane 3: VV1 50%/MM2 50%; Lane 4: VV1 25%/MM2 75%; Lane 5: MM2 100%. (G, H) Same mixtures were tested in the PK resistance ELISA assay. (G) VV2 100% (filled circles), VV2 75%/MM1 25% (filled triangles), VV2 50%/MM1 50% (filled inverted triangles), VV2 25%/MM1 75% (open triangles), MM1 100% (open circles). (H) MM2 100% (filled circles), MM2 75%/VV1 25% (filled triangles), MM2 50%/VV1 50% (filled inverted triangles), MM2 25%/VV1 75% (open triangles), VV1 100% (open circles).

Mentions: In sCJD and iCJD patients who harboured a single WB PrPSc type in the different brain areas, as assessed by Sha31, a single ELISA PK resistance profile (Table 1 and Figure 3A) and a comparable ratio in strain typing assay (Table 1 and Figure 3B) were observed in all brain areas. Surprisingly, in each patient harbouring both type 1 and 2 PrPres, either in the same or in different brain areas, a single ELISA PK digestion profile (Table 1 and Figure 3C and 3D) and a comparable signal ratio in strain typing assay (Table 1 and Figure 3B) was also observed, irrespective of region assayed.


Beyond PrP9res) type 1/type 2 dichotomy in Creutzfeldt-Jakob disease.

Uro-Coste E, Cassard H, Simon S, Lugan S, Bilheude JM, Perret-Liaudet A, Ironside JW, Haik S, Basset-Leobon C, Lacroux C, Peoch' K, Streichenberger N, Langeveld J, Head MW, Grassi J, Hauw JJ, Schelcher F, Delisle MB, Andréoletti O - PLoS Pathog. (2008)

PrPsc PK Resistance and Molecular Strain Variations in sCJD and iCJD Brain Samples.Each investigated brain sample was initially characterized by WB using antibody Sha31. Symbol patterns represent type 1 in white, type 1+2 in grey and type 2 in black. (A) Results from PK resistance ELISA carried out on three different brain areas (cerebellum, caudate nucleus and temporal cortex) from a MM1 (open circles), a VV1 (open triangles), a VV2 (inverted filled triangle) and a MM2 (filled squares) sCJD patient. Values obtained are expressed as percentage of signal obtained with the lowest PK concentration (50 µg/mL). (B) Results from CEA strain typing ELISA (one symbol per patient—3 to 5 different areas by patients). PrPSc signal intensity was measured after PK digestion into two different detergent solutions. Normalized A/A′ ratio was calculated for each sample (see Methods section). MM1 and MV1 had a low ratio indicating an absence of alteration of PrPsc PK sensitivity linked to the modification detergent digestion conditions. This ratio was higher in MV2, VV2, MV1+2, and VV1+2, while, in the unique VV1 case, an intermediate ratio was observed. In MM2 patients, the huge ratio indicated a strong increase in PK sensitivity by modification of detergent conditions. (C, D) PK resistance assay in three areas from a (C) VV (triangles) or MM (circles) patient and in (D) a MV (triangles) patient harbouring distinct PrPSc WB type in their different brain areas. Artificial mixtures of MM2/VV1 or MM1/VV2 samples were prepared. All homogenates were first equilibrated by dilution into negative brain homogenate to obtain an equal PrPSc signal in ELISA. (E, F) Mixtures were then tested by Western Blot (200 µg PK digestion—Sha31 anti PrP antibody). (E) Lane 1: MM1 100%; Lane 2: MM1 75%/VV2 25%; Lane 3: MM1 50%/VV2 50%; Lane 4: MM1 25%/VV2 75%; Lane 5: VV2 100%. (F) Lane 1: VV1 100%; Lane 2: VV1 75%/MM2 25%; Lane 3: VV1 50%/MM2 50%; Lane 4: VV1 25%/MM2 75%; Lane 5: MM2 100%. (G, H) Same mixtures were tested in the PK resistance ELISA assay. (G) VV2 100% (filled circles), VV2 75%/MM1 25% (filled triangles), VV2 50%/MM1 50% (filled inverted triangles), VV2 25%/MM1 75% (open triangles), MM1 100% (open circles). (H) MM2 100% (filled circles), MM2 75%/VV1 25% (filled triangles), MM2 50%/VV1 50% (filled inverted triangles), MM2 25%/VV1 75% (open triangles), VV1 100% (open circles).
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2279301&req=5

ppat-1000029-g003: PrPsc PK Resistance and Molecular Strain Variations in sCJD and iCJD Brain Samples.Each investigated brain sample was initially characterized by WB using antibody Sha31. Symbol patterns represent type 1 in white, type 1+2 in grey and type 2 in black. (A) Results from PK resistance ELISA carried out on three different brain areas (cerebellum, caudate nucleus and temporal cortex) from a MM1 (open circles), a VV1 (open triangles), a VV2 (inverted filled triangle) and a MM2 (filled squares) sCJD patient. Values obtained are expressed as percentage of signal obtained with the lowest PK concentration (50 µg/mL). (B) Results from CEA strain typing ELISA (one symbol per patient—3 to 5 different areas by patients). PrPSc signal intensity was measured after PK digestion into two different detergent solutions. Normalized A/A′ ratio was calculated for each sample (see Methods section). MM1 and MV1 had a low ratio indicating an absence of alteration of PrPsc PK sensitivity linked to the modification detergent digestion conditions. This ratio was higher in MV2, VV2, MV1+2, and VV1+2, while, in the unique VV1 case, an intermediate ratio was observed. In MM2 patients, the huge ratio indicated a strong increase in PK sensitivity by modification of detergent conditions. (C, D) PK resistance assay in three areas from a (C) VV (triangles) or MM (circles) patient and in (D) a MV (triangles) patient harbouring distinct PrPSc WB type in their different brain areas. Artificial mixtures of MM2/VV1 or MM1/VV2 samples were prepared. All homogenates were first equilibrated by dilution into negative brain homogenate to obtain an equal PrPSc signal in ELISA. (E, F) Mixtures were then tested by Western Blot (200 µg PK digestion—Sha31 anti PrP antibody). (E) Lane 1: MM1 100%; Lane 2: MM1 75%/VV2 25%; Lane 3: MM1 50%/VV2 50%; Lane 4: MM1 25%/VV2 75%; Lane 5: VV2 100%. (F) Lane 1: VV1 100%; Lane 2: VV1 75%/MM2 25%; Lane 3: VV1 50%/MM2 50%; Lane 4: VV1 25%/MM2 75%; Lane 5: MM2 100%. (G, H) Same mixtures were tested in the PK resistance ELISA assay. (G) VV2 100% (filled circles), VV2 75%/MM1 25% (filled triangles), VV2 50%/MM1 50% (filled inverted triangles), VV2 25%/MM1 75% (open triangles), MM1 100% (open circles). (H) MM2 100% (filled circles), MM2 75%/VV1 25% (filled triangles), MM2 50%/VV1 50% (filled inverted triangles), MM2 25%/VV1 75% (open triangles), VV1 100% (open circles).
Mentions: In sCJD and iCJD patients who harboured a single WB PrPSc type in the different brain areas, as assessed by Sha31, a single ELISA PK resistance profile (Table 1 and Figure 3A) and a comparable ratio in strain typing assay (Table 1 and Figure 3B) were observed in all brain areas. Surprisingly, in each patient harbouring both type 1 and 2 PrPres, either in the same or in different brain areas, a single ELISA PK digestion profile (Table 1 and Figure 3C and 3D) and a comparable signal ratio in strain typing assay (Table 1 and Figure 3B) was also observed, irrespective of region assayed.

Bottom Line: However, since cases of CJD show co-occurrence of type 1 and type 2 PrP(res) in the brain, the basis of this classification system and its relationship to agent strain are under discussion.In 30% of cases, both type 1 and type 2 PrP(res) were identified.Despite this, the other two biochemical assays found that PrP(Sc) from an individual patient demonstrated uniform biochemical properties.

View Article: PubMed Central - PubMed

Affiliation: INSERM U858, Institut de Médecine Moléculaire de Rangueil and Service d'Anatomie Pathologique et Histologie-Cytologie, C.H.U. Rangueil, Toulouse, France.

ABSTRACT
Sporadic Creutzfeldt-Jakob disease (sCJD) cases are currently subclassified according to the methionine/valine polymorphism at codon 129 of the PRNP gene and the proteinase K (PK) digested abnormal prion protein (PrP(res)) identified on Western blotting (type 1 or type 2). These biochemically distinct PrP(res) types have been considered to represent potential distinct prion strains. However, since cases of CJD show co-occurrence of type 1 and type 2 PrP(res) in the brain, the basis of this classification system and its relationship to agent strain are under discussion. Different brain areas from 41 sCJD and 12 iatrogenic CJD (iCJD) cases were investigated, using Western blotting for PrP(res) and two other biochemical assays reflecting the behaviour of the disease-associated form of the prion protein (PrP(Sc)) under variable PK digestion conditions. In 30% of cases, both type 1 and type 2 PrP(res) were identified. Despite this, the other two biochemical assays found that PrP(Sc) from an individual patient demonstrated uniform biochemical properties. Moreover, in sCJD, four distinct biochemical PrP(Sc) subgroups were identified that correlated with the current sCJD clinico-pathological classification. In iCJD, four similar biochemical clusters were observed, but these did not correlate to any particular PRNP 129 polymorphism or western blot PrP(res) pattern. The identification of four different PrP(Sc) biochemical subgroups in sCJD and iCJD, irrespective of the PRNP polymorphism at codon 129 and the PrP(res) isoform provides an alternative biochemical definition of PrP(Sc) diversity and new insight in the perception of Human TSE agents variability.

Show MeSH
Related in: MedlinePlus